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Vol. 11, Issue 3, 969-982, March 2000
§ and
*Section of Microbiology, University of California, Davis,
California 95616; The cytoplasm-to-vacuole targeting (Cvt) pathway and macroautophagy
are dynamic events involving the rearrangement of membrane to form a
sequestering vesicle in the cytosol, which subsequently delivers its
cargo to the vacuole. This process requires the concerted action of
various proteins, including Apg5p. Recently, it was shown that another
protein required for the import of aminopeptidase I
(API) and autophagy, Apg12p, is covalently attached to Apg5p through
the action of an E1-like enzyme, Apg7p. We have undertaken an
analysis of Apg5p function to gain a better understanding of the role
of this novel nonubiquitin conjugation reaction in these import
pathways. We have generated the first temperature-sensitive mutant in
the Cvt pathway, designated apg5ts.
Biochemical analysis of API import in the
apg5ts strain confirmed that Apg5p is
directly required for the import of API via the Cvt pathway. By
analyzing the stage of API import that is blocked in the
apg5ts mutant, we have determined that Apg5p
is involved in the sequestration step and is required for vesicle
formation and/or completion.
Department of Chemical and Biological
Sciences, Faculty of Science, Japan Women's University, Mejirodai,
Tokyo 112, Japan; Department of Cell Biology, National
Institute for Basic Biology, Okazaki 444-8585, Japan; and
§Precursory Research for Embryonic Science and Technology,
Japan Science and Technology Corporation, Kawaguchi 332-0012, Japan
Corresponding author. E-mail address:
djklionsky{at}ucdavis.edu.
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